Presence Of 1-hydroxybenzotriazole Research Articles

Oxidation of carbazole, N-ethylcarbazole, fluorene, and dibenzothiophene by the laccase of Coriolopsis gallica

Purified laccase from Coriolopsis gallica UAMH8260 oxidized carbazole, N-ethylcarbazole, fluorene, and dibenzothiophene in the presence of 1-hydroxybenzotriazole and 2,2′-azinobis (3-ethylbenzthiazoline-6-sulfonic acid) as free radical mediators. Susceptibility to laccase oxidation appears related to the ionization potential (IP) of the substrate: compounds with an IP above 8.52, dibenzofuran (IP = 8.77) and benzothiophene (IP = 8.73) were not attacked. Carbazole (IP = 7.68) was the most sensitive to oxidation with >99% transformed with 10 milliunits of laccase after 1 h, though most reactions were carried out for 18 h. 9-Fluorenone was identified as the product of fluorene (IP = 8.52) oxidation, and dibenzothiophene sulfone from dibenzothiophene (IP = 8.44). Although carbazole and N-ethylcarbazole were both completely removed within 18 h, no oxidation or condensation metabolites were detected. This investigation is the first to report the oxidation of dibenzothiophene, carbazole, and N-ethylcarbazole by laccase.

Aromatic ring cleavage of a non-phenolic β- O-4 lignin model dimer by laccase of Trametes versicolor in the presence of 1-hydroxybenzotriazole

The novel cleavage products, 2,3-dihydroxy-1-(4-ethoxy-3-methoxyphenyl)-1-formyloxypropane (II) and 1-(4-ethoxy-3-methoxyphenyl)-1,2,3-trihydroxypropane-2,3-cyclic carbonate (III) were identified as products of a non-phenolic β- O-4 lignin model dimer, 1,3-dihydroxy-2-(2,6-dimethoxylphenoxy)-1-(4-ethoxy-3-methoxyphenyl)propane (I), by a Trametes versicolor laccase in the presence of 1-hydroxybenzotriazole (1-HBT). An isotopic experiment with a 13C-labeled lignin model dimer, 1,3-dihydroxy-2-(2,6-[U-ring- 13C]dimethoxyphenoxy)-1-(4-ethoxy-3-methoxyphenyl)propane (I- 13C) indicated that the formyl and carbonate carbons of products II and III were derived from the β-phenoxy group of β- O-4 lignin model dimer I as aromatic ring cleavage fragments. These results show that the laccase-1-HBT couple could catalyze the aromatic ring cleavage of non-phenolic β- O-4 lignin model dimer in addition to the β-ether cleavage, Cα-Cβ cleavage, and Cα-oxidation.

Synthesis of Ordered Polymer by Direct Polycondensation. 9. Ordered Poly(amide−acylhydrazide−amide) from Three Nonsymmetric Monomers

An ordered (−ccfebaddabef−) poly(amide−acylhydrazide−amide) was prepared by the direct polycondensation of 5-(3-carboxypropyl)-2-isopropoxybenzoic acid (XabX), bis(4-nitrophenyl) isophthalate (XccX), piperazine (YddY), and 4-aminobenzohydrazide (YefY), using the condensing agent diphenyl(2,3-dihydro-2-thioxo-3-benzoxazolyl)phosphonate (1). The polymerization was carried out by mixing the dicarboxylic acid, diester, condensing agent 1, and triethylamine (TEA) in N-methyl-2-pyrrolidinone (NMP) for 3 h at room temperature, followed by adding piperazine and aminobenzohydrazide at −15 °C. The mixture was stirred at −15 °C for 2 h, room temperature for 24 h, and 70 °C for 4 d in the presence of 1-hydroxybenzotriazole (HOBt). The resulting polymer gave an inherent viscosity of 0.20 dL/g in NMP, measured at a concentration of 0.5 g/dL at 30 °C. The feasibility of the formation of the ordered polymer from three nonsymmetric monomers was demonstrated by studying the model reactions in detail. Furthermore, the authentic ordered and random polymers were prepared to verify the structure of the ordered polymer. The microstructures of the polymers obtained were investigated by 1H and 13C NMR spectroscopy, and it was found that the polymer obtained by direct polycondensation has the expected ordered structure.

Oxidation of Phenanthrene by a Fungal Laccase in the Presence of 1-Hydroxybenzotriazole and Unsaturated Lipids

Phenanthrene, a polycyclic aromatic hydrocarbon, was efficiently oxidized by laccase in the presence of both 1-hydroxybenzotriazole and unsaturated lipids. 73 % of initially added phenanthrene was degraded within 182 hours to give phenanthrene-9,10-quinone and 2,2′-diphenic acid as the major products. The system was also able to peroxidize linoleic acid to its corresponding hydroperoxides suggesting the involvement of lipid peroxidation in laccase catalyzed phenanthrene oxidation. Lipid peroxidation by laccase required 1-hydroxybenzotriazole and did not depend on Mn2+and H2O2suggesting that the chemical reactions involved differ from those previously reported for manganese peroxidase.

Studies on sensitivity to racemization of activated residues in couplings of N-benzyloxycarbonyldipeptides.

A series of 24 peptides Z-Gly-Xaa(R)-OH where Xaa = 15 different residues and R = H, NH2, tBu, Bzl, Trt, Mtr, and StBu were coupled with valine benzyl ester in dimethylformamide or dichloromethane at +5 degrees. The accompanying racemization was determined by analysis of the epimeric products by normal phase high-performance liquid chromatography (HPLC) for Xaa(R) = Met, Cys(StBu) and Lys(Z) and by reversed-phase HPLC after removal of benzyl-based protecting groups for Xaa(R) = Ser(tBu), Thr(tBu) and Arg(Mtr). The coupling methods examined included mixed anhydride (MxAn) at -5 degrees, and N,N'-dicyclohexylcarbodiimide (DCC), benzotriazol-1-yl-tris(dimethylamino)phosphonium hexafluorophosphate (BOP) and O-benzotriazol-1-yl-N,N,N',N'-tetramethyluroniumhexafluorophosp hate (HBTU) in the presence of 1-hydroxybenzotriazole (HOBt). Very few couplings gave stereochemically pure products. The order of sensitivity to racemization of residues depended on the method of coupling and the solvent. It varied most when comparing MxAn to HOBt-assisted reactions; it varied moderately when comparing HOBt-assisted reactions. There was less variation in comparing BOP and HBTU reactions that are initiated by the same mechanism. Leu, Nle, Phe, Asn, Lys(Z) and Asp(OBzl) are identified as the residues least sensitive to racemization. DCC-HOBt generally led to less epimerization than the other methods.

Fmoc-amino acid chlorides in solid phase synthesis of opioid peptides.

Fmoc-amino acid chlorides were employed in the solid phase synthesis of the opioid peptides [Leu]enkephalin, [Leu]enkephalin amide, and dermorphin. The conventional polystyrene-based Merrifield resin or Wang's resin served as solid support. A binary salt of either triethylamine or diisopropylethylamine in the presence of 1-hydroxybenzotriazole or pivalic acid was used for acylation. The coupling rates were quite fast, being comparatively faster when 1-hydroxybenzotriazole was used along with triethylamine or diisopropylethylamine. The peptides obtained in good yields showed, after purification, biological and spectral properties identical with those of the natural peptides.

Fluorinated amino acids and peptides. Synthesis of 3,3-difluoro-2-amino acids, peptides and cyclodipeptides incorporating 3,3-difluoro-2-aminobutyric acid or 3,3-difluorophenylalanine residues in their structures

Convenient syntheses have been developed for N-9-fluorenylmethyloxycarbonyl-DL-2-amino-3,3-difluoro-butanoic acid and N-9-fluorenylmethyloxycarbonyl-DL-3,3-difluorophenylalanine which have been successfully introduced into peptide syntheses. Several fluorinated peptides have been obtained either by active ester coupling methods, mixed anhydride synthesis or with the aid of dicyclohexylcarbodiimide (DCC) in presence of 1-hydroxybenzotriazole (HOBt). The method has been applied to the preparation of seven hitherto unpublished representative 3-(1,1′-difluoroalkyl)-2,5-dioxopiperazines for biological purposes and structural chemistry studies.

Orthogonal solid‐phase synthesis of human gastrin‐I under mild conditions*

An approach to the solid-phase segment condensation synthesis of the 17-peptide amide human gastrin-I has been developed. N alpha-amino and side-chain protection were provided by 9-fluorenylmethyloxycarbonyl (Fmoc) and tert.-butyl groups, and a series of anchors cleavable under mild conditions were used. The N-terminal pentapeptide pGlu-Gly-Pro-Trp-Leu-OH was prepared using a p-alkoxybenzyl ester linkage made by a preformed handle strategy. Cleavage, in 65% yield, was with the new Reagent M: CF3COOH-CH2Cl2-beta-mercaptoethanol--anisole (70:30:2:1), which was optimized to preserve the labile tryptophan residue. A new preformed handle procedure expedited solid-phase synthesis of the protected "middle" hexapeptide, Fmoc-(Glu(OtBu]5-Ala-OH, anchored as an o-nitrobenzyl ester. Chains were not lost during this assembly, and final photolytic cleavage (350 nm) in toluene--CF3CH2OH (4:1) occurred in 59% yield. Both protected intermediates were purified by simple gel filtration, whereupon they were shown to be pure by analytical HPLC, and gave satisfactory NMR and FABMS spectra. Last, the C-terminal hexapeptide, Tyr(tBu)-Gly-Trp-Met-Asp(OtBu)-Phe, was assembled on a tris(alkoxy)benzylamide "PAL" support. For the polymer-supported segment condensation, the middle and N-terminal pieces were added respectively in greater than 98% and 89% yields (judged by amino acid analysis and solid-phase sequencing), by overnight couplings in N,N-dimethylformamide (DMF) mediated by benzotriazolyl N-oxytrisdimethylaminophosphonium hexafluorophosphate (BOP) in the presence of 1-hydroxybenzotriazole (HOBt) and N-methylmorpholine (NMM). Racemization was 4% and 11% respectively at Ala and Leu. Cleavage with Reagent M followed by reversed-phase chromatography gave pure gastrin-I in an overall 30% isolated yield. These results compare favorably with those from a stepwise assembly.

Improved solid phase synthesis of luteinizing hormone releasing hormone analogues using 9-fluorenylmethyloxycarbonyl amino acid active esters and catalytic transfer hydrogenation with minimal side-chain protection and their biological activities

Using mainly 9-fluorenylmethyloxycarbonyl amino acid 2, 4, 5-trichlorophenyl esters in the presence of 1-hydroxybenzotriazole and the solid supportp-alkoxybenzyl alcohol resin, synthesis of luteinizing hormone releasing hormone analogues was carried out with minimal side-chain protection. Catalytic transfer hydrogenation was employed for removal of NO2 and Z-groups from Arg and < Glu respectively avoiding the use of HF and this led to good yields. An aromatic, hydrophilic amino acid, D-(p-hydroxyphenyl) glycine was incorporated into luteinizing hormone releasing hormone molecule along with other modifications. The agonistic as well as antagonistic activities of all the peptides have been studied

New analogues of leucine-methionine-enkephalin

Nine analogues of the opioid pentapeptides leucine-/ methionine-enkephalinamide, involving replacement of amino acid at position 5 or amino acids at positions 2 and 5, have been synthesized by the solid phase method using mainly 9-fluorenylmethyloxycarbonyl amino acid trichlorophenyl esters in the presence of 1-hydroxybenzotriazole, the solid support being the Merrifield resin. All the analogues were effective in inhibiting the electri cally stimulated contractions of the guinea pig ileum (in vitro) and one of them, tyrosyl-Dnorvalyl-glycyl-phenylalanyl-methioninamide was found to be about 82 times more active than morphine. They also exhibited analgesic activity as well as antidiarrhoeal activity in mice (in vivo).

λ -Benzyl-L-Glutamate Graft Copolymers of Cellulose and Poly[Arylene Ether Sulfone

Abstract Macroinitiators with primary amino substituents were synthesized by one of the following techniques: a) cyanoethylation of cellulose followed by diborane reduction to produce aminopropylcellulose, 1; b) nitration, then SnCl2 reduction of poly(arylene ether sulfone), 5, to produce poly(2-aminoarylene ether sulfone), 2; c) phthalimidation of 5 followed by hydrazinolysis to yield poly(2-aminomethylarylene ether sulfone), 3; and d) LiAlH4 reduction of poly(cyanophenylene arylene ether) to poly(aminomethylphenylene arylene ether), 4. Heterogeneous grafting of Λ-benzyl-L-glutamate-N-carboxyanhydride, 8, to Polymer 1 resulted in a nonrandom distribution of amino acid residues; α-helical conformations were detected at low BLG-NCA/NH2 ratios (<5 amino acids). Using molar ratios ranging from 1 to 100 of 8, relative to the amine concentration, grafting to Polymers 3 and 4 was effected in anhydrous THF at room temperature under homogeneous conditions. If reaction times between 24 and 48 h are utilized, high grafting efficiencies (>80%) are obtained. The conformation of the polypeptide chain was evaluated by NMR and infrared spectroscopy. Polypeptides grafted to Polymers 3 and 4 appeared to adopt the expected conformation for the chain length predicted, i.e., a progression from random coil (<8 amino acids) to β-pleated sheet (8–13 amino acids) to α-helix (> 13 amino acids). The benzyl ester functions on the BLG grafts are subject to direct modification with amine nucleophiles; studies with butylamine correlate reaction conditions with extent of ester vs peptide cleavage. In the presence of 1-hydroxybenzotriazole, aminolysis of the ester is favored and conversions to Λ-amides up to 75% without peptide cleavage are achieved.

ChemInform Abstract: Synthesis of Modified Tuftsins Containing Monosaccharides or Monosaccharide Derivatives.

Synthesis of some modified tuftsins is described in which a monosaccharide or a monosaccharide derivative was incorporated in the molecule. Acylation of H-Thr-Lys(Z)-Pro-Arg(NO2)-OBzl with D(+)-gluco-1,5-lactone followed by catalytic hydrogenation gave N alpha-gluconyl-tuftsin. Glycosylation of the carboxyl function of the C-terminal arginine has been achieved by reacting, through the mixed anhydride procedure, Boc-Thr-Lys(Z)-Pro-OH with 2-deoxy-2-(NG-nitroargininamido)-D-glucopyranose followed by catalytic hydrogenation and trifluoroacetic acid treatment. O-Glucosyl-tuftsin has been prepared by reacting o-nitrophenyl N-benzyloxycarbonyl-O-[(alpha + beta) 2,3,4,6-tetra-O-benzyl-D-glucopyranosyl]-threoninate with H-Lys(Z)-Pro-Arg(NO2)-OBzl in the presence of 1-hydroxybenzotriazole. Flash chromatography on silica gel allowed a partial separation of the diastereoisomers, one of which has been isolated in a reasonable yield. The single diastereoisomer and the alpha + beta anomeric mixture were separately deblocked by catalytic hydrogenation and purified by RP-HPLC.

Human plasma apolipoprotein C-II: total solid-phase synthesis and chemical and biological characterization.

The complete amino acid sequence of human plasma apolipoprotein C-II (apoC-II) has been synthesized chemically by the solid-phase method using phenylacetamidomethyl-resin. All amino acids were coupled to the peptide-resin in the presence of 1-hydroxybenzotriazole; tert-butyloxycarbonyl-protected amino acids with the appropriate side-chain-protecting groups that are stable to the reaction conditions used in the solid-phase methodology were used. After cleavage and deprotection, the crude apoC-II was purified by ion-exchange chromatography and then by reverse-phase high-performance liquid chromatography. The purified apolipoprotein was found to elute as a single peak under various chromatographic conditions, and the overall yield of the final purified protein was 20.7%. Synthetic apoC-II was characterized by several complementary analytical techniques including amino acid composition, Edman phenylisothiocyanate degradation, polyacrylamide gel electrophoresis, and high-performance liquid chromatography. The final product was found to be homogeneous and to activate normal human post-heparin lipase to the same extent as native apoC-II. The synthetic protein is also equally immunoreactive as native apoC-II.

Synthesis of modified tuftsins containing monosaccharides or monosaccharide derivatives.

Synthesis of some modified tuftsins is described in which a monosaccharide or a monosaccharide derivative was incorporated in the molecule. Acylation of H-Thr-Lys(Z)-Pro-Arg(NO2)-OBzl with D(+)-gluco-1,5-lactone followed by catalytic hydrogenation gave N alpha-gluconyl-tuftsin. Glycosylation of the carboxyl function of the C-terminal arginine has been achieved by reacting, through the mixed anhydride procedure, Boc-Thr-Lys(Z)-Pro-OH with 2-deoxy-2-(NG-nitroargininamido)-D-glucopyranose followed by catalytic hydrogenation and trifluoroacetic acid treatment. O-Glucosyl-tuftsin has been prepared by reacting o-nitrophenyl N-benzyloxycarbonyl-O-[(alpha + beta) 2,3,4,6-tetra-O-benzyl-D-glucopyranosyl]-threoninate with H-Lys(Z)-Pro-Arg(NO2)-OBzl in the presence of 1-hydroxybenzotriazole. Flash chromatography on silica gel allowed a partial separation of the diastereoisomers, one of which has been isolated in a reasonable yield. The single diastereoisomer and the alpha + beta anomeric mixture were separately deblocked by catalytic hydrogenation and purified by RP-HPLC.

Synthesis and biological activity of some new leucine-enkephalin analogues

The opioid pentapeptide leucine-enkephalinamide and eleven of its analogues have been synthesised by the solid phase technique employing mostly 9-fluoroenylmethyloxycarbonyl amino acid active esters in the presence of 1-hydroxybenzotriazole. Both the conventional chloromethylated copolystyrene-2% divinylbenzene resin as well asp-alkoxybenzyl alcohol resin were employed and it was observed that yields were uniformly better with the latter resin. The analogues were made by affecting single or multiple replacements of amino acids involving positions 1,2 and 5. Some of the analogues were found to be more potent than morphine in the guinea pig ileum assay.

Semisynthetic derivatives of glucagon. The contribution of histidine-1 to hormone conformation and activity.

Semisynthetic N epsilon- acetimidoglucagon was prepared from the [des- His1 ]analogue by coupling the N-hydroxysuccinimide ester of N alpha- tBoc - Nimidazole -DNP-L-histidine to the peptide in dimethylformamide in the presence of 1-hydroxybenzotriazole. The deprotected, purified product was chemically identical to N epsilon- acetimidoglucagon and equipotent to N epsilon- acetimidoglucagon and native glucagon in its ability to activate adenylate cyclase and displace [125I] iodoglucagon from rat liver plasma membranes. Semisynthetic [ Phe1 ]-, [ Ala1 ]-, and [des- His1 ] glucagons prepared similarly achieved 85, 55, and 35% of the maximal activity and 22, 2, and 6% of the binding potency of N epsilon- acetimidoglucagon . The biological assays indicate that the amino group is involved to a greater extent in transduction than in binding, but the aromatic nature and hydrogen bonding capability of the imidazole ring of histidine-1 are important for both binding and transduction. In circular dichroism studies, all derivatives exhibited increased helicity in 2-chloroethanol. The [ Phe1 ] analogue although less soluble behaved similarly to native glucagon, while the [ Ala1 ] and [des- His1 ] derivatives exhibited an increased helical content in 0.01 N HCl as a result of an increased propensity of these derivatives to self-associate in the absence of 2-chloroethanol. The unexpected conformational changes throughout the molecule may have relevance for the functional activity.

Chemical synthesis of urotensin II, a somatostatin like peptide in the caudal neurosecretory system of fishes.

In the goby, Gillichthys mirabilis, urotensin II (a bioactive neuropeptide present in the urophysis of teleost fish) has the dodecapeptide sequence, H2N-AGTADC-FWKYCV-OH, which is homologous with mammalian somatostatin at positions 1, 2 and 7-9. The Merrifield solid phase synthesis of Gillichthys urotensin II (UII) was accomplished by stepwise assembly from the carboxy terminus using N-alpha-tert, butyloxycarbonyl (Boc) amino acids containing benzyl-derived groups for protection of side-chain functionalities, Coupling of amino acids to the growing peptide was mediated by diisopropylcarbodiimide (DIC) in the presence of 1-hydroxybenzotriazole (HOBt). Residual alpha-amino groups remaining after coupling were blocked by acetylation with 1-acetylimidazole. Crude, synthetic UII was extracted from the HF-treated, protected peptide-resin product, reduced with dithiothreitol (DTT), reoxidized at high dilution with O2, and separated into its components using a single, preparative, reverse-phase HPLC step. The pure, synthetic UII, obtained in 7.6% yield from oxidized crude UII, was indistinguishable from pure, native UII in specific bioactivity, amino acid sequence, and retention time in each of two different HPLC systems.

SYNTHESIS OF NUCLEOSIDE PHOSPHOTRIESTERS CONTAINING AMIDATE OR THIOATE FUNCTIONS

Abstract Phosphorylation of properly protected nucleosides with aryl or alkyl phosphorodichloridates, in the presence of 1-hydroxybenzotriazole, gives phosphorylated intermediates which react smoothly with different amine or thiol functions to afford the corresponding amidate or thioate phosphotriester derivatives.

Synthesis of human beta-endorphin in solution using benzyl-type side chain protective groups.

A solution synthesis of human beta-endorphin (beta-EP) was carried out by condensation of protected peptide segments bearing N alpha-tert.-butyloxycarbonyl groups and benzyl-derived groups for the protection of functionalities in amino acid side chains. Five intermediate segments were assembled in a stepwise manner starting at the carboxyl terminus. Thus, the segment of sequence region (27-31) was coupled to segment (22-26) by the azide method. Segment (19-21) was incorporated into the growing chain by azide coupling, and segment (10-18) by dicyclohexylcarbodiimide coupling in the presence of 1-hydroxybenzotriazole (DDC-HOBt). Solubility problems in condensing the ensuing 22-peptide with segment (1-9) by DDC-HOBt were overcome by using a dimethylformamidephenol mixture as a solvent. Protecting group cleavage by Na in liquid NH3 was much superior to liquid HF which gave rise to many decomposition products. Homogeneous betah-EP indistinguishable from authentic material in physiochemical and biological properties, was obtained in a single preparative reversed phase liquid chromatographic step after protecting group cleavage.

ChemInform Abstract: SOLID‐PHASE SYNTHESIS OF OXYTOCIN, DESAMINOOXYTOCIN AND 4‐THR‐OXYTOCIN USING ACTIVE ESTERS IN PRESENCE OF 1‐HYDROXYBENZOTRIAZOLE

Chemischer InformationsdienstVolume 10, Issue 7 Natural Products ChemInform Abstract: SOLID-PHASE SYNTHESIS OF OXYTOCIN, DESAMINOOXYTOCIN AND 4-THR-OXYTOCIN USING ACTIVE ESTERS IN PRESENCE OF 1-HYDROXYBENZOTRIAZOLE S. A. KHAN, S. A. KHANSearch for more papers by this authorK. M. SIVANANDAIAH, K. M. SIVANANDAIAHSearch for more papers by this author S. A. KHAN, S. A. KHANSearch for more papers by this authorK. M. SIVANANDAIAH, K. M. SIVANANDAIAHSearch for more papers by this author First published: February 13, 1979 https://doi.org/10.1002/chin.197907305AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume10, Issue7February 13, 1979 RelatedInformation